LOCUS       sequence-140194-        7040 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7040)
  AUTHORS   Itakura E, Kishi-Itakura C, Mizushima N
  TITLE     The hairpin-type tail-anchored SNARE syntaxin 17 targets to 
            autophagosomes for fusion with endosomes/lysosomes.
  JOURNAL   Cell. 2012 Dec 7;151(6):1256-69. doi: 10.1016/j.cell.2012.11.001.
  PUBMED    23217709
REFERENCE   2  (bases 1 to 7040)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2012.11"; journalName: "Cell"; date: "2012-12-7- 7";
            volume: "151"; issue: "6"; pages: "1256-69"
COMMENT     Sequence Label: FLAG-SNAP29
FEATURES             Location/Qualifiers
     source          1..7040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             614..679
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1482..2104
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1681..1700)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2128..2148)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2146..2443
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2497..3291
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2551..2570)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3161..3180
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3943..4077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3980..3999)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4034..4053
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4082..4101)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4083..4101)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4115..4131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4115..4131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4128..4150)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4139..4155
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4163..4169,4170..4187,4188..4193))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4208..4229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4346..4363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4517..5105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4597..4616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5276..6067,6068..6136))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5899..5918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6137..6241)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6268..6723
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6355..6374)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6565..6586
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6849..6871
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6863..6880
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6864..6880
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattaacc atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta
      661 caaggatgac gatgacaagc ttgcggccgc gaattccatg tcagcttacc ctaaaagcta
      721 caatccgttc gacgacgacg gggaggacga aggcgcccgg ccggcccctt ggagggacgc
      781 ccgagacctc cccgacgggc ccgacgcgcc cgcggacagg cagcagtact tgcggcagga
      841 ggtcctccgc agggctgagg ccacggccgc cagcaccagc aggtccctgg ccctcatgta
      901 cgagtccgag aaggttgggg tcgcctcttc cgaggagctc gcccgtcagc gaggagtcct
      961 ggagcgcaca gagaagatgg tggacaagat ggaccaagat ttgaagatca gccagaaaca
     1021 catcaatagc attaagagcg tgtttggggg gctggtcaat tacttcaaat ccaaaccagt
     1081 agagacccca cctgaacaga atggcaccct cacctcccag cccaacaaca gattgaaaga
     1141 agctataagt acaagtaaag aacaggaagc aaagtaccag gccagccacc caaaccttag
     1201 aaagctggat gatacagacc ctgtccccag aggggctggt tctgccatga gtactgatgc
     1261 ttacccaaag aacccacacc ttcgagccta tcaccagaag atcgacagca acctagatga
     1321 gctgtccatg ggactgggtc gtctgaagga catagccctg gggatgcaga cagaaattga
     1381 ggagcaagat gacattcttg accggctgac aaccaaagtg gacaagttag atgtcaacat
     1441 aaaaagcaca gaaagaaaag ttcgacaact ctgaggatcc cgggtggcat ccctgtgacc
     1501 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     1561 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     1621 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     1681 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     1741 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     1801 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     1861 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     1921 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc
     1981 agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt
     2041 tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg
     2101 aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2161 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2221 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2281 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2341 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2401 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggaatt gatcagcttg
     2461 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     2521 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     2581 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     2641 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     2701 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     2761 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     2821 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     2881 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     2941 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3001 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3061 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3121 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3181 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3241 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     3301 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     3361 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     3421 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccgggctc gatcccctcg
     3481 cgagttggtt cagctgctgc ctgaggctgg acgacctcgc ggagttctac cggcagtgca
     3541 aatccgtcgg catccaggaa accagcagcg gctatccgcg catccatgcc cccgaactgc
     3601 aggagtgggg aggcacgatg gccgctttgg tcgacccgga cgggacgctc ctgcgcctga
     3661 tacagaacga attgcttgca ggcatctcat gagtgtgtct tcccgttttc cgcctgaggt
     3721 cactgcgtgg atggagcgct ggcgcctgct gcgcgacggc gagctgctca ccacccactc
     3781 gccaagctgg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgccgatc
     3841 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3901 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3961 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4021 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaat
     4081 tccctatagt gagtcgtatt aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt
     4141 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4201 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4261 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4321 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4381 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4441 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4501 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4561 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4621 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4681 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4741 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4801 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4861 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4921 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4981 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5041 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5101 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5161 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5221 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5281 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5341 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5401 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5461 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5521 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5581 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5641 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5701 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5761 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5821 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5881 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5941 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6001 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6061 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6121 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6181 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6241 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     6301 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     6361 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     6421 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     6481 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     6541 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     6601 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     6661 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     6721 tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     6781 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     6841 agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctga tctatacatt
     6901 gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg
     6961 ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg
     7021 tccaatatga ccgccatgtt
//
